Genotypic and Phenotypic Characterization of Bacteriocinogenic Lactic Acid Bacterial Strains for Possible Beneficial, Virulence, and Antibiotic Resistance Traits

Abstract

Lactic acid bacteria (LAB) are a diverse group of microorganisms that can be found in natural habitats and are used in a wide range of industries, especially in the food sector. They are regarded "Generally Recognized as Safe" and are commonly used as a starter culture, probiotics, and biopreservative. However, some LAB genera are excluded from the "Qualified Presumption of Safety" (QPS) list due to the possibility that they have virulence and antibiotic resistance genetic determinants. The objective of this study was to investigate the beneficial, virulence, and antibiotic resistance-related genes in 10 well-characterized bacteriocinogenic LAB strains. The auto-aggregation, co-aggregation, cell surface hydrophobicity, and bile salt tolerance were also evaluated to examine their potential as bacteriocinogenic probiotics and/or starter culture. Results showed that all tested strains showed abilities for auto-aggregation at 4oC and 37oC, co-aggregation with S. aureus JCM8704, S. typhimurium BIOTECH1826, and E. coli DH5α, and significant cell-surface hydrophobicity. However, only a few strains were able to withstand the media treated with 0.3% bile salt. Among the tested bacteriocinogenic LAB strains, L. lactis IO-1 and C. divergens V41 had the maximum values for auto-aggregation at 4oC and 37oC, respectively. C. divergens V41 also exhibited the highest percentage for cell surface hydrophobicity. E. faecium NKR-5-3 showed the highest co-aggregation percentages with all indicator strains. Our findings also showed that the tested isolates presented distinct combinations of virulence-related genes. Only two of ten bacteriocinogenic LAB strains exhibited presence of multiple virulence genes. Lactococcus sp. QU12 was found to have a high frequency of beneficial and virulence genes, with 2 out of 7 genes present encoding beneficial factor and 11 out of 13 genes encoding virulence factor. Lactococcus sp. QU12 and L. lactis IO-1 were also positive for tetracycline resistance gene tetM and aminoglycoside resistance gene aphA-2, respectively, and transposon genes. Moreover, only a few LAB isolates tested positive for 2 out of 8 antibiotic resistance classes. Although few, the substantial danger of these genes being transferred and acquired cannot be overlooked as this could potentially cause serious health concerns in the future. These results suggest that despite the promising properties of bacteriocinogenic LAB, careful safety evaluation of these strains should be a prerequisite before using these in food systems.